title: Isolation and molecular analysis of genes Stpk-V2 and Stpk-V3 homologous to powdery mildew resistance gene Stpk-V in a Dasypyrum villosum accession and its derivatives.
abstract: Wheat-Dasypyrum villosum translocated chromosomes T6V#2S•6AL and T6V#4S•6DL are known to confer excellent resistance to wheat powdery mildew (PM). However, it is difficult to distinguish the two sources of PM resistance genes through multi-pathotype testing because to date no virulence for them has been found. To reveal the relationship between the PM resistance genes from the two translocations, the sequence of the Stpk-V gene, a key member of powdery mildew resistance locus Pm21, was used as a reference to isolate homologous genes from a D. villosum accession No.1026 and its derivatives 6V#4(6D) disomic substitution (DS) line RW15 and T6V#4S•6DL translocation line Pm97033. Two genes Stpk-V2 and Stpk-V3 were cloned from No.1026. Sequence alignment showed that Stpk-V2 and Stpk-V3 shared 98.2 % and 96.2 % of their DNA and 99.3 % and 100 % of their amino acids in identity with Stpk-V. Compared with Stpk-V, a 22-bp direct sequence repeat and a miniature inverted-repeat transposable element (MITE) were found in the intron 4 of Stpk-V2 and Stpk-V3, respectively. However, Stpk-V2 was not present in DS line RW15 and translocation line Pm97033 based on the PCR result, indicating that Stpk-V2 did not contribute to the PM resistance of RW15 and Pm97033. In the promoter region, a 78-bp insertion was found not only in Stpk-V2 and Stpk-V3, but also in its orthologous gene Stpk-A of wheat. In addition, there was a 17 bp/8 bp deletion/insertion in the putative promoter of Stpk-V3 in comparison with that of Stpk-V/Stpk-V2. Real-time quantitative RT-PCR analysis indicated that the expression levels of Stpk-V and Stpk-V3 genes in the translocation lines were induced by the pathogen, but Stpk-V had a higher expression level than Stpk-V3 at 12 h after inoculation with Bgt. The diversity of Stpk-V gene will help to explore new resistance genes to PM in D. villosum for wheat breeding.
authors.affiliation: National Key Facility for Crop Gene Resources and Genetic Improvement / Key Laboratory of Biology and Genetic Improvement of Triticeae Crops, Ministry of Agriculture / Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing, 100081, China.
authors.forename: Z S
authors.initials: ZS
authors.lastname: Lin
authors.forename: Y L
authors.initials: YL
authors.lastname: Zhang
authors.forename: M J
authors.initials: MJ
authors.lastname: Wang
authors.forename: J R
authors.initials: JR
authors.lastname: Li
authors.forename: K
authors.initials: K
authors.lastname: Wang
authors.forename: X
authors.initials: X
authors.lastname: Chen
authors.forename: Q F
authors.initials: QF
authors.lastname: Xu
authors.forename: X S
authors.initials: XS
authors.lastname: Zhang
authors.forename: X G
authors.initials: XG
authors.lastname: Ye
mesh_terms: D000595:Amino Acid Sequence; D001203:Ascomycota / Q000472:pathogenicity*; D001483:Base Sequence; D018744:DNA, Plant / Q000235:genetics; D060467:Disease Resistance / Q000235:genetics*; D017343:Genes, Plant; D008969:Molecular Sequence Data; D010935:Plant Diseases / Q000382:microbiology*; D006109:Poaceae / Q000235:genetics*; D011401:Promoter Regions, Genetic; D018749:RNA, Plant / Q000235:genetics; D017422:Sequence Analysis, DNA; D014178:Translocation, Genetic
chemical_list: D018744:DNA, Plant; D018749:RNA, Plant